In Vitro Anticancer Activity on 3T3, PC3, and HeLa Cell Line and GC-MS Assay of Oil Fractions of Alstonia Scholaris Flower Obtained by Column Chromatography

Kaneez Fatima, Shaukat Khalid, Kiran Qadeer, Hina Yasin, Hina Abrar, Raheela Bano, Adeel Arsalan, Rana Asif Hussain

Abstract

Several plant-derived compounds are currently successfully employed in cancer treatment. Various studies have demonstrated the anticancer or cytotoxic potential of different extracts of Alstonia scholaris at the highest doses. This study aimed to analyze the anticancer potential of oil fractions obtained from Alstonia scholaris flower isolation. The oil fractions were characterized using gas chromatography–mass spectrometry (GC-MS) assay. The (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide) (MTT) colorimetric assay was used for 3T3, PC3, and HeLa cell line cytotoxic potential of oil fractions. The results showed no cytotoxic activity of oil fractions, i.e., Oil I and Oil II were at a minimum dose of 30 μg/ml. However, the GC-MS assay depicted saturated (palmitic acid, ethyl ester, caproic acid, stearic acid, ethyl ester, octacosanoic acid, etc.), unsaturated (oleic acid, 3-(octadecyloxy)propyl ester, ethyl oleate and some other hydrocarbons such as betulin, ethyl iso-allocholate, lupenyl acetate, phthalic anhydride, phytol, etc. Thus, the reported chemical constituents showed no significant anticancer effect in the scientific literature. This research suggested that Alstonia has no potential as an anticancer agent. The findings of this study provide insight into maintaining the optimum data for future aspects of plant-derived phytoconstituents with potential cytotoxic effects on 3T3, PC3, and HeLa cell lines.   

 

Keywords: Alstonia scholaris, GC-MS assay, MTT colorimetric assay, cytotoxicity.

 

https://doi.org/10.55463/issn.1674-2974.49.11.2


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CHANDA S., and NAGANI K. In vitro and in vivo methods for anticancer activity evaluation and some Indian medicinal plants possessing anticancer properties: an overview. Journal of Pharmacognosy and Phytochemistry, 2013, 2(2): 140-152. https://www.phytojournal.com/archives/2013/vol2issue2/PartB/20.1.pdf

JAHAN S., CHAUDHARY R., and GOYAL P.K. Anticancer activity of an Indian medicinal plant, Alstonia scholaris, on skin carcinogenesis in mice. Integrative cancer therapies, 2009, 8(3): 273-279. https://doi.org/10.1177/1534735409343590

CIAPETTI G., CENNI E., PRATELLI L., and PIZZOFERRATO A. In vitro evaluation of cell/biomaterial interaction by MTT assay. Biomaterials, 1993, 14(5): 359-364. https://doi.org/10.1016/0142-9612(93)90055-7

KUMAR P., NAGARAJAN A., and UCHIL P.D. Analysis of cell viability by the MTT assay. Cold Spring Harbor Protocols, 2018, 6. DOI:10.1101/pdb.prot095497

GHAFFAR N.I.A. Synthesis and Biological Evaluation of Novel Analogues of the Plant Derived Natural Products, Incarviditone and Incarvilleatone. PhD Thesis. The University of Manchester, UK, 2020. https://www.proquest.com/openview/20e959be217a62422f9ab553659d507a/1?pq-origsite=gscholar&cbl=2026366&diss=y

VAN MEERLOO, J., KASPERS G.J.L., and CLOOS J. Cell sensitivity assays: the MTT assay. Methods in Molecular Biology, 2011, 731: 237-245. DOI: 10.1007/978-1-61779-080-5_20.

AMADOR M.L, JIMENO J., PAZ-ARES L., CORTES-FUNES H., and HIDALGO M. Progress in the development and acquisition of anticancer agents from marine sources. Annals of Oncology, 2003, 14 (21): 1607-1615. DOI: https://doi.org/10.1093/annonc/mdg443

BHAKUNI D.S., BITTNER M., MARTICORENA C., SILVA M., WELDT E., and HOENEISEN M. Screening of Chilean plants for anticancer activity. I. Lloydia, 1976, 39(4): 225-243. https://europepmc.org/article/med/957912

KHAZIR J., RILEY D.L., PILCHER L.A., DE-MAAYER P., and MIR B.A. Anticancer agents from diverse natural sources. Natural Product Communications, 2014, 9(11): 1655-1669. DOI: 1934578X1400901130. https://doi.org/10.1177/1934578X1400901130

RAINA H., SONI G., JAUHARI N., SHARMA N., and BHARADVAJA N. Phytochemical importance of medicinal plants as potential sources of anticancer agents. Turkish Journal of Botany, 2014, 38(6): 1027-1035. DOI: 10.3906/bot-1405-93

SHAH U., SHAH R., ACHARYA S., and ACHARYA N. Novel anticancer agents from plant sources. Chinese Journal of Natural Medicines, 2013, 11(1): 16-23. https://doi.org/10.1016/S1875-5364(13)60002-3

SOLOWEY E., LICHTENSTEIN M., SALLON S., PAAVILAINEN H., SOLOWEY E., and LORBERBOUM-GALSKI H. Evaluating medicinal plants for anticancer activity. The Scientific World Journal, 2014, 721402. DOI: 10.1155/2014/721402

FOUCHÉ G., CRAGG G.M., PILLAY P., KOLESNIKOVA N., MAHARAJ V.J., and SENABE J. In vitro anticancer screening of South African plants. Journal of Ethnopharmacology, 2008, 119(3): 455-461.

GAIDHANI S.N., SINGH A., KUMARI S., LAVEKAR G.S., JUVEKAR A.S., SEN S., and PADHI M.M. Evaluation of some plant extracts for standardization and anticancer activity. Indian Journal of Traditional Knowledge, 2013, 12(4): 682-687.

SAKLANI A., and KUTTY S.K. Plant-Derived Compounds In Clinical Trials. Drug Discovery Today, 2008, 13(3-4): 161-171. https://doi.org/10.1016/j.jep.2008.07.005

BALIGA M.S. Alstonia scholaris Linn R Br in the treatment and prevention of cancer: past, present, and future. Integrative Cancer Therapies. 2010, 9(3): 261-269. DOI: 10.1177/1534735410376068

KAUSHIK P., KAUSHIK D., SHARMA N., and RANA A.C. Alstonia scholaris: It's phytochemistry and pharmacology. Chronicles of Young Scientists, 2011, 2(2). link.gale.com/apps/doc/A261829166/AONE?u=anon~605b8b70&sid=googleScholar&xid=06640a68.

ADOTEY J.P.K., ADUKPO G.E., OPOKU BOAHEN Y., and ARMAH F.A. A review of the ethnobotany and pharmacological importance of Alstonia boonei De Wild (Apocynaceae). ISRN Pharmacology, 2012, Volume 2012, Article ID 587160. DOI:10.5402/2012/587160

JAHAN N., SUBRIN S., and HAQUE M.R. In vitro Study of Different Partitionates of Alstonia scholaris (L.) R. Br. Leaf for Thrombolytic and Membrane Stabilizing Activities. Bangladesh Pharmaceutical Journal, 2018, 21(2): 145-149. https://doi.org/10.3329/bpj.v21i2.37926

ARULMOZHI S., MAZUMDER P.M., ASHOK P., and NARAYANAN L.S. Pharmacological activities of Alstonia scholaris Linn. (Apocynaceae)-A review. Pharmacognosy Reviews, 2007, 1(1): 163-170. http://www.phcogrev.com/article/2007/1/1-18?qt-sidebar_tabs=0

FATIMA K., KHALID S., QADEER K., YASIN H., ARSALAN A., ABRAR H., ZAHID S., HUSSAIN R.A., ISLAM M., and ALI M.S. Urease inhibition and DPPH radical scavenging potential of phytoconstituent from Alstonia scholaris and molecular docking interactions of bioactive luteolin with target proteins. Pakistan Journal of Pharmaceutical Sciences, 2022, 35(1): 219-225. https://web.p.ebscohost.com/abstract?direct=true&profile=ehost&scope=site&authtype=crawler&jrnl=1011601X&AN=155540985

GREEN H., and KEHINDE O. An established preadipose cell line and its differentiation in culture II. Factors affecting the adipose conversion. Cell, 1975, 5(1):19-27. https://doi.org/10.1016/0092-8674(75)90087-2

LI L., WANG H., YANG E.S., ARTEAGA C.L., and XIA F. Erlotinib attenuates homologous recombinational repair of chromosomal breaks in human breast cancer cells. Cancer Research, 2008, 68(22): 9141-9146. https://doi.org/10.1158/0008-5472.CAN-08-1127

TAI S., SUN Y., SQUIRES J.M., ZHANG H., OH W.K., LIANG C.Z., and HUANG J. PC3 is a cell line characteristic of prostatic small cell carcinoma. The Prostate, 201, 71(15): 1668-1679. https://doi.org/10.1002/pros.21383

GHAFFARI S.R., SABOKBAR T., MOLLAHAJIAN H., DASTAN J., RAMEZANZADEH F., ENSANI F., YARANDI F., MOUSAVI-JARRAHI A., MOHAGHEGHI M.A., and MORADI A. Prevalence of human papillomavirus genotypes in women with normal and abnormal cervical cytology in Iran. Asian Pacific Journal of Cancer Prevention, 2006, 7(4): 529-532. http://www.scopus.com/inward/record.url?eid=2-s2.0...

BÖTTCHER M., and BECK O. Evaluation of buprenorphine CEDIA assay versus GC-MS and ELISA using urine samples from patients in substitution treatment. Journal of Analytical Toxicology, 2005, 29(8): 769-776. https://doi.org/10.1093/jat/29.8.769

LIU W., MORROW J.D., and YIN H. Quantification of F2-isoprostanes as a reliable index of oxidative stress in vivo using gas chromatography–mass spectrometry (GC-MS) method. Free Radical Biology and Medicine. 2009, 47(8): 1101-1107. https://doi.org/10.1016/j.freeradbiomed.2009.07.028

DĄBROWSKI Ł. Evaluation of a Simplified Method for GC/MS Qualitative Analysis of Polycyclic Aromatic Hydrocarbons, Polychlorinated Biphenyls, and Organic Pesticides Using PARADISe Computer Program. Molecules, 2020, 25(16): 3727. https://doi.org/10.3390/molecules25163727

JAGETIA G.C., and BALIGA M.S. Effect of Alstonia scholaris in enhancing the anticancer activity of berberine in the Ehrlich ascites carcinoma-bearing mice. Journal of Medicinal Food, 2004, 7(2): 235-244. DOI: 10.1089/1096620041224094

JAGETIA G.C., and BALIGA M.S. Evaluation of anticancer activity of the alkaloid fraction of Alstonia scholaris (Sapthaparna) in vitro and in vivo. Phytotherapy Research, 2006, 20(2): 103-109. https://doi.org/10.1002/ptr.1810

JAHAN S., CHAUDHARY R., and GOYAL P.K. Anticancer activity of an Indian medicinal plant, Alstonia scholaris, on skin carcinogenesis in mice. Integrative Cancer Therapies, 2009, 8(3): 273-279. doi/abs/10.1177/1534735409343590

JAGETIA G.C., and BALIGA M.S. The effect of seasonal variation on the antineoplastic activity of Alstonia scholaris R. Br. in HeLa cells. Journal of Ethnopharmacology, 2005, 96(1-2): 37-42. https://doi.org/10.1016/j.jep.2004.07.024

PRASANNARAJ G., and VENKATACHALAM P. Green engineering of biomolecule-coated metallic silver nanoparticles and their potential cytotoxic activity against cancer cell lines. Advances in Natural Sciences: Nanoscience and Nanotechnology, 2017, 8(2): 025001.

BADYAL A., SHARMA V., KOUR N., and SINGH S.K. Anticancer efficacy of methanolic extracts of some medicinal plants from Jammu region, Jammu & Kashmir, India. Indian Journal of Biochemistry & Biophysics, 2016, 53(1): 51-56.

VAGHORA B., and SHUKLA V. Impact of different phytochemical classes and Ayurvedic plants in battle against cancer. Skin, 2016, 13: 14.

ZURAIDA, and MARIYA S. In vitro cytotoxicity of Alstonia scholaris (R. Br) bark on Vero and HeLa cell lines. IOP Conference Series: Earth and Environmental Science, 2019, 374, 012065. https://iopscience.iop.org/article/10.1088/1755-1315/374/1/012065/meta

MOSMANN T. Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assays. Journal of Immunological Methods, 1983, 65: 55-63.


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